上皮间质转化相关蛋白在肝细胞肝癌组织中的表达鉴定及其小分子RNA表达谱的研究

目的 检测E-钙黏蛋白(E-cadherin)和波形蛋白(vimentin)及其相应小分子RNA(miRNA)在原发性肝细胞肝癌(HCC)中的表达.方法 免疫组织化学检测32例HCC组织中E-cadherin和vimentin的表达,分析其与临床病理资料的关系.应用miRNA芯片筛选HCC转移相关差异表达miRNAs.选取部分差异表达miRNAs以实时定量逆转录-聚合酶链反应(RT-PCR)方法进行验证,利用在线靶基因预测软件对其靶基因进行预测.结果 E-cadherin在转移、低分化/未分化、大于3 cm组中的阳性表达率分别为25.0%(2/8)、35.7%(5/14)、52.9%(9/17),vimentin在上述三组的阳性表达率分别为87.5%(7/8)、71.4%(10/14)、52.9%(9/17).E-cadherin表达下调、vimentin表达上调与HCC分化(低分化/未分化)和转移明显相关(P＜0.05),而与其大小无关.miRNA芯片筛选获得36个HCC转移相关miRNAs(8个上调,28个下调).在E-cadherin(-)/vimentin(+)转移HCC中,miR-21、miR-221的表达(2.22±1.79、2.36±1.05)明显高于E-cadherin (+)/vimentin(-)无转移HCC(4.35±1.90、3.90±1.23,P＜0.05);miR-199a、miR-126的表达(4.42±0.61、3.62±0.54)明显低于E-cadherin(+)/vimentin(-)无转移HCC(2.43±0.85、2.54±1.10,P＜0.05).结论 E-cadherin(-)/vimentin(+)转移HCC与E-cadherin(+)/vimentin(-)无转移HCC的miRNA明显差异表达,其中部分差异表达miRNAs可能通过调控上皮间质转化相关分子的表达参与HCC的转移.

Abstract：

Objective To detect the expression of E-cadherin and vimentin, and their corresponding microRNAs (miRNAs) in primary heptocellular carcinoma (HCC). Methods The expression of E-cadherin and vimentin in 32 cases of HCC tissues was examined by using immunohistochemistry, and the relationship between the expression and clinicopathology was analyzed. miRNA array was used to investigate the differentially expressed miRNAs between E-cadherin ( - )/vimentin ( + ) metastasis HCC and E-cadherin ( + )/vimentin ( - ) no-metastasis HCC. Real-time polymerase chain reaction (PCR) was applied to verify the reliability of miRNA array results. We predicted target genes of the four miRNAs by combination anticipated algorithms. Results The positive rate of E-cadherin in the metastasis, poor/no differentiation, ＞3 cm groups was 25.0% (2/8), 35.7% (5/14), 52. 9% (9/17) respectively, and that of vimentin was 87.5% (7/8), 71.4% (10/14), 52.9% (9/17)respectively. The low expression of E-cadherin and high expression of vimentin in HCC was significantly correlated with metastasis and poor differentiation of HCC (P＜0. 05). miRNA array showed that 8 miRNAs were up-regulated and 28 miRNAs were down-regulated in E-cadherin ( - )/vimentin ( + ) metastasis HCC. The expression levels of miR-21 and miR-221 in E-cadherin ( - )/vimentin ( + ) metastasis HCC were 2. 22 ± 1.79 and 2. 36 ±1.05, significantly higher than those in E-cadherin ( + )/vimentin ( - ) no-metastasis HCC (4. 35 ±1.90, 3.90 ± 1.23,P＜0.05). The expression levels of miR-126 and miR-199a in E-cadherin (-)/vimentin ( + ) metastasis HCC were 4. 42 ± 0. 61 and 3.62 ± 0. 54, notably lower than those in E-cadherin ( +)/vimentin (-) no-metastasis HCC (2.43±0.85, 2.54±-1.10,P＜0.05). Conclusion miRNA differential expression profile of E-cadherin ( - )/vimentin ( + ) HCC was obtained, and some of miRNAs may play a role in metastasis of HCC by regulating epithelial to mesenchymal transition.

The expression detection of epithelial to mesenchymal transition related proteins and screening of its differential expression microRNAs in hepatocellular carcinoma

Objective To detect the expression of E-cadherin and vimentin, and their corresponding microRNAs (miRNAs) in primary heptocellular carcinoma (HCC). Methods The expression of E-cadherin and vimentin in 32 cases of HCC tissues was examined by using immunohistochemistry, and the relationship between the expression and clinicopathology was analyzed. miRNA array was used to investigate the differentially expressed miRNAs between E-cadherin ( - )/vimentin ( + ) metastasis HCC and E-cadherin ( + )/vimentin ( - ) no-metastasis HCC. Real-time polymerase chain reaction (PCR) was applied to verify the reliability of miRNA array results. We predicted target genes of the four miRNAs by combination anticipated algorithms. Results The positive rate of E-cadherin in the metastasis, poor/no differentiation, ＞3 cm groups was 25.0% (2/8), 35.7% (5/14), 52. 9% (9/17) respectively, and that of vimentin was 87.5% (7/8), 71.4% (10/14), 52.9% (9/17)respectively. The low expression of E-cadherin and high expression of vimentin in HCC was significantly correlated with metastasis and poor differentiation of HCC (P＜0. 05). miRNA array showed that 8 miRNAs were up-regulated and 28 miRNAs were down-regulated in E-cadherin ( - )/vimentin ( + ) metastasis HCC. The expression levels of miR-21 and miR-221 in E-cadherin ( - )/vimentin ( + ) metastasis HCC were 2. 22 ± 1.79 and 2. 36 ±1.05, significantly higher than those in E-cadherin ( + )/vimentin ( - ) no-metastasis HCC (4. 35 ±1.90, 3.90 ± 1.23,P＜0.05). The expression levels of miR-126 and miR-199a in E-cadherin (-)/vimentin ( + ) metastasis HCC were 4. 42 ± 0. 61 and 3.62 ± 0. 54, notably lower than those in E-cadherin ( +)/vimentin (-) no-metastasis HCC (2.43±0.85, 2.54±-1.10,P＜0.05). Conclusion miRNA differential expression profile of E-cadherin ( - )/vimentin ( + ) HCC was obtained, and some of miRNAs may play a role in metastasis of HCC by regulating epithelial to mesenchymal transition.